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Perkinsus Medium

山東拓普生物工程有限公司 培養(yǎng)基配方 http://www.jlkcpj.cn

Composition per liter:

NaCl..............................................................................................9.0g
NaHCO 3 ........................................................................................2.1g
Glucose .........................................................................................1.1g
NaH 2 PO 4 ·H 2 O.............................................................................0.29g
KCl..............................................................................................0.38g
L- Arginine·HCl............................................................................0.21g
L- Glutamine.................................................................................0.30g
MgSO 4 ·7H 2 O..............................................................................0.17g
Sodium pyruvate.........................................................................0.11g
KH 2 PO 4 .......................................................................................0.08g
CaCl 2 ·2H 2 O.................................................................................0.09g
L- Cystine·2HCl............................................................................0.04g
L- Lysine·HCl...............................................................................0.04g
L- Leucine.......................................................................................0.2g
L- Isoleucine.................................................................................0.02g
L- Histidine·HCl·H 2 O...................................................................0.02g
L- Arginine ...................................................................................0.02g
L- Threonine.................................................................................0.02g
L- Valine .......................................................................................0.02g
L- Tyrosine....................................................................................0.02g
L- Methionine............................................................................0.02mg
L- Cystine ...................................................................................0.016g
L- Phenylalanine.........................................................................0.014g
L- Serine .......................................................................................0.01g
L- Asparagine·H 2 O.......................................................................0.01g
L- Aspartic Acid...........................................................................0.01g
L- Glutamic acid...........................................................................0.01g
L- Histidine...................................................................................0.01g
L- Proline......................................................................................0.01g
L- Glycine.....................................................................................0.01g
L- Alanine....................................................................................8.9mg
D- Phenylalanine..........................................................................5.0mg
L- Methionine..............................................................................4.5mg
Hypoxanthine.............................................................................4.1mg
L- Tryptophan..............................................................................4.6mg
L- Threonine................................................................................3.6mg
L- Valine ..................................................................................... 3.5mg
L- Tyrosine...................................................................................1.8mg
Vitamin B 12 ................................................................................1.4mg
Folic acid....................................................................................2.3mg
Phenol Red.................................................................................1.2mg
Thiamine·HCl ............................................................................2.0mg
FeSO 4 ·7H 2 O...............................................................................0.8mg
Choline chloride.........................................................................1.7mg
Calcium  DL -pantothenate...........................................................1.7mg
Thymidine..................................................................................0.7mg
Niacinamide...............................................................................1.6mg
Pyridoxal....................................................................................1.0mg
Inositol.......................................................................................0.7mg
Riboflavin ..................................................................................0.5mg
Lipoic acid .................................................................................0.2mg
Pyridoxine·HCl..........................................................................0.2mg
ZnSO 4 ·7H 2 O............................................................................0.03mg
FeNO 3 ·7H 2 O..........................................................................0.025mg
Biotin .......................................................................................0.02mg
CuSO 4 ·5H 2 O...............................................................................3.0μg
SASW solution ......................................................................910.0mL
HEPES (N-[2-hydroxyethyl] piperazine-
N′-2-ethanesulfonic acid) buffer (1.0M solution) .............25.0mL
Fetal bovine serum, heat inactivated .......................................20.0mL
JLP carbohydrate solution.......................................................10.0mL
Lipid concentrate (100X).........................................................10.0mL
NaHCO 3  solution.......................................................................8.6mL
L -Glutamine solution.................................................................5.0mL
Phenol Red solution...................................................................0.5mL
SASW Solution:
Composition per liter:
Seawater, synthetic basal mixture...............................................18.2g
Preparation of SASW Solution: Add seawater synthetic basal
mixture to distilled/deionized water and bring volume to 1.0L. Mix
thoroughly.
JLP Carbohydrate Solution:
Composition per 100.0mL:
Glucose.........................................................................................5.0g
Galactose.......................................................................................1.0g
Trehalose.......................................................................................1.0g
Preparation of JLP Carbohydrate Solution: Add components
to distilled/deionized water and bring volume to 100.0mL. Mix thor-
oughly.
Phenol Red Solution:
Composition per 10.0mL:
Phenol Red..................................................................................0.05g
Preparation of Phenol Red Solution:  Add Phenol Red to dis-
tilled/deionized water and bring volume to 10.0mL.
Glutamine Solution:
Composition per 10.0mL:
L -Glutamine................................................................................0.29g
Preparation of Glutamine Solution:  Add  L -glutamine to dis-
tilled/deionized water and bring volume to 10.0mL.
NaHCO 3 Solution:
Composition per 10.0mL:
NaHCO 3 ......................................................................................0.75g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/de-
ionized water and bring volume to 10.0mL.
Lipid Concentrate:
Composition per liter:
Pluronic™ F68............................................................................10.0g
Tween™ 80...................................................................................2.5g
Cod liver oil..................................................................................1.0g
Cholesterol..................................................................................0.45g
DL -α-Tocopherol acetate...............................................................0.2g
Preparation of Lipid Concentrate: Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Filter ster-
ilize.
Preparation of Medium: Add all components, except lipid concen-
trate and fetal bovine serum, to distilled/deionized water and bring vol-
ume to 1.0L. Mix thoroughly. Filter sterilize. Aseptically add 20.0mL
of sterile fetal bovine serum and 10.0mL sterile lipid concentrate. Mix
thoroughly. Aseptically distribute into sterile tubes or flasks. Use im-
mediately.
Use: For the cultivation of Perkinsus marinus, P. andrewsi, P. chesa-
peaki, and P. atlanticus.